Fig. 3: Regulation of ptsG′-′lacZ translational fusion by SgrS point mutants.

a Regulation of chromosomal ptsG′-′lacZ translational fusion by wild-type SgrS and A177U, G178U, G178A, G184A, G215A, U224A, and U224G mutant variants (plasmid-encoded) was assessed using β-galactosidase activity assay. Data were obtained from n = 4 independent experiments and presented as mean ± SEM. b RNA was extracted simultaneously with β-galactosidase activity assay and northern blot was performed on two biological replicates using probes specific for SgrS sRNA and 5S rRNA (control). Full-length (227 nt), properly terminated SgrS transcripts are labeled as “termination” products, and longer transcripts that arose due to transcriptional readthrough are labeled as “readthrough” products. c Band intensities of total SgrS transcripts (termination + readthrough) were measured, and 5S-normalized values were plotted as “SgrS abundance” (steady-state transcript abundance of SgrS mutants). Data were obtained from n = 2 independent experiments and presented as mean ± SEM. d Band intensities of SgrS termination and readthrough products were measured and 5S-normalized ratios (readthrough/termination) were calculated and plotted for each SgrS mutant as “SgrS readthrough ratio”. Data were obtained from n = 2 independent experiments and presented as mean ± SEM. Source data are provided as a Source data file.